Tracing MYC expression for small molecule discovery
Files
Accepted manuscript
Date
2019-05-16
Authors
Steinberger, Jutta
Robert, Francis
Hallé, Maxime
Williams, David E.
Cencic, Regina
Sawhney, Neha
Pelletier, Dylan
Williams, Philip
Igarashi, Yasuhiro
Version
Accepted manuscript
OA Version
Citation
Jutta Steinberger, Francis Robert, Maxime Hallé, David E Williams, Regina Cencic, Neha Sawhney, Dylan Pelletier, Philip Williams, Yasuhiro Igarashi, John A Porco, Abimael D Rodriguez, Brigitte Kopp, Brian Bachmann, Raymond J Andersen, Jerry Pelletier. 2019. "Tracing MYC expression for small molecule discovery." Cell Chem Biol, Volume 26, Issue 5, pp. 699 - 710.e6. https://doi.org/10.1016/j.chembiol.2019.02.007
Abstract
Our inability to effectively "drug" targets such as MYC for therapeutic purposes requires the development of new approaches. We report on the implementation of a phenotype-based assay for monitoring MYC expression in multiple myeloma cells. The open reading frame (ORF) encoding an unstable variant of GFP was engineered immediately downstream of the MYC ORF using CRISPR/Cas9, resulting in co-expression of both proteins from the endogenous MYC locus. Using fluorescence readout as a surrogate for MYC expression, we implemented a pilot screen in which ∼10,000 compounds were prosecuted. Among known MYC expression inhibitors, we identified cardiac glycosides and cytoskeletal disruptors to be quite potent. We demonstrate the power of CRISPR/Cas9 engineering in establishing phenotype-based assays to identify gene expression modulators.